Cytochrome c oxidase, the terminal enzyme of the mitochondrial respiratory chain, catalyzes the electron transfer from reduced cytochrome c to oxygen. It is a heteromeric complex consisting of 3 catalytic subunits encoded by mitochondrial genes and multiple structural subunits encoded by nuclear genes. The mitochondrially-encoded subunits function in electron transfer, and the nuclear-encoded subunits may be involved in the regulation and assembly of the complex. This nuclear gene encodes subunit VIc, which has 77% amino acid sequence identity with mouse subunit VIc. This gene is up-regulated in prostate cancer cells. A pseudogene has been found on chromosomes 16p12.

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Western blotting analysis using COX6C antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with COX6C antibody and HRP-conjugated goat anti-mouse secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Western blotting analysis using COX6C antibody . COX6C expression in wild-type (WT) and COX6C shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with COX6C antibody and HRP-conjugated goat anti-mouse secondary antibody respectively. Image was developed using NaQ™ ECL Substrate Kit .

Immunocytochemical staining of HAP-1 cells with COX6C antibody . Nuclei were stained blue with DAPI; COX6C was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Flow cytometric analysis of COX6C expression in HAP-1 cells using COX6C antibody . Green, isotype control; red, COX6C.

Validation of COX6C knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with COX6C antibody and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using COX6C antibody , Top panel: wild-type (WT); Bottom panal: COX6C shRNA knockdown (KD). Nuclei were stained blue with DAPI; Acyl-CoA dehydrogenase short chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

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